Our goals with this dataset were to 1) isolate, culture, and identify two fungal life stages of Aspergillus flavus, 2) characterize the volatile emissions from grain inoculated by each fungal morphotype, and 3) understand how microbially-produced volatile organic compounds (MVOCs) from each fungal morphotype affect foraging, attraction, and preference by S. oryzae. This dataset includes that derived from headspace collection coupled with GC-MS, where we found the sexual life stage of A. flavus had the most unique emissions of MVOCs compared to the other semiochemical treatments. This translated to a higher arrestment with kernels containing grain with the A. flavus sexual life stage, as well as a higher cumulative time spent in those zones by S. oryzae in a video-tracking assay in comparison to the asexual life stage. While fungal cues were important for foraging at close-range, the release-recapture assay indicated that grain volatiles were more important for attraction at longer distances. There was no significant preference between grain and MVOCs in a four-way olfactometer, but methodological limitations in this assay prevent broad interpretation. Overall, this study enhances our understanding of how fungal cues affect the foraging ecology of a primary stored product insect. In the assays described herein, we analyzed the behavioral response of Sitophilus oryzae to five different blends of semiochemicals found and introduced in wheat (Table 1). Briefly, these included no stimuli (negative control), UV-sanitized grain, clean grain from storage (unmanipulated, positive control), as well as grain from storage inoculated with fungal morphotype 1 (M1, identified as the asexual life stage of Aspergillus flavus) and fungal morphotype 2 (M2, identified as the sexual life stage of A. flavus). Fresh samples of semiochemicals were used for each day of testing for each assay. In order to prevent cross-contamination, 300 g of grain (tempered to 15% grain moisture) was initially sanitized using UV for 20 min. This procedure was done before inoculating grain with either morphotype 1 or 2. The 300 g of grain was kept in a sanitized mason jar (8.5 D × 17 cm H). To inoculate grain with the two different morphologies, we scraped an entire isolation from a petri dish into the 300 g of grain. Each isolation was ~1 week old and completely colonized by the given morphotype. After inoculation, each treatment was placed in an environmental chamber (136VL, Percival Instruments, Perry, IA, USA) set at constant conditions (30°C, 65% RH, and 14:10 L:D). This procedure was the same for both morphologies and was done every 2 weeks to ensure fresh treatments for each experimental assay. See file list for descriptions of each data file. Resources in this dataset:Resource Title: Ethovision Movement Assay. File Name: ponce_lizarraga_ethovision_assay_microbial_volatiles_2020.csvResource Software Recommended: Excel,url: https://www.microsoft.com/en-us/microsoft-365/excel Resource Title: Olfactometer Round 1 Assay - With Fused Air Permeable Glass. File Name: ponce_lizarraga_first_round_olfactometer_fungal_study_2020.csvResource Software Recommended: Excel,url: https://www.microsoft.com/en-us/microsoft-365/excel Resource Title: Olfactometer Round 2 Assay - With Fused Air Permeable Glass Containing Holes. File Name: ponce_lizarraga_second_round_olfactometer_fungal_study_2021.csvResource Software Recommended: Excel,url: https://www.microsoft.com/en-us/microsoft-365/excel Resource Title: Small Release-Recapture Assay. File Name: ponce_lizarraga_small_release_recapture_assay.csvResource Software Recommended: Excel,url: https://www.microsoft.com/en-us/microsoft-365/excel Resource Title: Large Release-Recapture Assay. File Name: ponce_lizarraga_large_release_recapture_assay.csvResource Software Recommended: Excel,url: https://www.microsoft.com/en-us/microsoft-365/excel Resource Title: Headspace Volatile Collection Assay. File Name: sandra_headspace_volatiles_2020.csvResource Software Recommended: Excel,url: https://www.microsoft.com/en-us/microsoft-365/excel Resource Title: README file list. File Name: file_list_stored_grain_Aspergillus_Sitophilus_oryzae.txt